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Objectives: To determine hepatitis B virus (HBV) genotypes in southern Seville (Spain) and investigate the development of lamivudine-resistance mutations by using a hybridization technique with specific probes and by comparing the results with those of the direct sequencing technique. To evaluate the temporal relationship between variations in the level of HBV-DNA and detection of mutant variants. To analyze the influence of several genotypes on the pattern of mutations developed and on values of viral load and alanine aminotransferase (ALT) after their development., Patients and Method: In 37 patients with chronic HBV infection, HBV genotype was determined using the LiPA technique. In 10 of these patients undergoing lamivudine treatment for a mean of 19.2 months, the development of lamivudine-resistant mutations was investigated. In these 10 patients, the LiPA technique was compared with direct sequencing. During lamivudine treatment, we determined HBV-DNA by polymerase chain reaction (PCR) and ALT every 3-6 months., Results: The most frequent genotypes were D (45.9%) and A (18.9%); 2 patients were genotype B while 18.9% had mixed genotypes. Sequencing showed identical results except in one mixed genotype. Mutations were found in 60% of the cases. The results of sequencing were in agreement, except in the detection of mixed populations composed of mutants and wild-type (WT). Patients with genotype A showed the pattern M204I+WT in the first 12 months and those with genotype D showed the pattern L180M+M204V with or without WT at 18 months. In 5/6 cases, an increase of > 1 log10 in HBV-DNA was observed 3-8 months before the mutation was detected by LiPA. In patients with genotype B, levels of HBV-DNA and ALT after the development of mutations was lower than basal levels and was also lower than those in patients with genotypes A and D., Conclusions: The LiPA technique for determination of HBV genotype and detection of lamivudine-resistance mutations shows excellent correlation with the most complex sequencing technique. Genotype D predominates in southern Seville. During lamivudine treatment, an increase in the level of HBV-DNA detected by PCR predicts the development of mutations before these are demonstrated by LiPA.

Background: There is scarce information about the influence of pregnancy in patients with chronic hepatitis C virus infection is little know., Patients and Methods: 6,556 pregnant women were screened for anti-HCV (ELISA II). We determine ALT, HCV-RNA by PCR (Amplicor Roche) and HCV viraemia (Amplicor-HCV-Monitor Roche) in the third trimester of pregnancy and after 6 months of delivery. HBsAg, anti-HIV and HCV serotype (Murex 1-3) were also determined., Statistical Analysis: Fisher test, paired-t and U Mann Whitney., Results: Anti-HCV was positive in 59 out of 6,556 (0.9%). Mean (SD) age: 27 (9) years (range, 18-40). Drug users: 34 (57%), post-transfusion: 10 (18%) and unknown: 15 (25%). HIV positive 11 (19%). Serotype 1, 30 (51%), setotype 3, 7 (20%), and nontypeable, 22 (37%). We studied HCV-RNA before and after delivery in 35 women, 8 out of 35 (23%) had HCV-RNA negative in both analysis. ALT was normal in 88% of women during pregnancy and in 42% after delivery. ALT levels in pregnancy were 32.6 (39.5) and in postpartum 64.5 (53.4) U/l (p < 0.005). 6 women were RNA-VHC negative during pregnancy and positive in postpartum. HCV viraemia during pregnancy and postpartum was 503 (1,203) and 1,014 (1,907) thousand copies/ml (p < 0.05). No relation was found among ALT or HCV viraemia with risk factors, serotype or coinfection with HIV., Conclusions: The prevalence of anti-HCV in pregnant women is 0.9%. ALT is usually normal in pregnancy. A quarter of women were HCV-RNA negative in pregnancy and positive after delivery. The viraemia was lower in pregnancy than after delivery, which is consistent with the fact of the low mother-to-infant HCV transmission rate.

The objective of this study was to evaluate the usefulness of blood culture in the diagnosis of disseminated mycobacteria (DMB). This prospective study included all blood cultures done for patients with fever and under suspicion of having DMB between January 1991 and July 1992. Fifty-seven blood samples from 16 patients were cultured; 14 (87.5%) patients were HIV positive and all were diagnosed as having DMB. The cultures were processed by lysis-centrifugation and identification of mycobacteria was by hybridization with a DNA probe. Mycobacterial growth was detected in 5 cultures (8.7%) from 4 patients (25%) (3 HIV positive). M. tuberculosis was isolated in 3 and M. avium in 1. Mean time until isolation was 46 days. In all cases mycobacteria were isolated in other samples before they were found in cultures: M. tuberculosis was isolated in 2 bronchial aspirates (BAS), 2 in liver tissue (L), 2 in spleen tissue (S), one in alveolar bronchial lavage, one in sputum, one in spinal fluid (SF) and one in urine. M. avium was isolated in sputum and ALB. The three patients in whom M. tuberculosis was found died 1.4 and 32 days after admission. In samples from the 12 DMB patients with negative cultures (11 HIV positive, 92%), M. tuberculosis was isolated in 100% of ganglion and S samples, 90% in urine, 69% in sputum, 67% in ABL and LB, 63% in BAS and 33% in SF. None of these patients died in hospital. We find blood culture to be of little use in the diagnosis of DMB. Analysis of other samples leads to faster diagnosis.(ABSTRACT TRUNCATED AT 250 WORDS)

We studied the sensibility to several antimicrobics of aeromonas strains isolated in extraintestinal infections during a period of five years, in order to establish which could be the best empirical therapy in these processes. Twenty-two strains of A. hydrophila were evaluated, was isolated (8 from hemocultures, 6 from wound exudate, 2 from abscess aspirate, 2 from peritoneal fluid and 1 from urine, pleural fluid, bile and catheter). The identification was made using the GNI card System (Vitek System) and other tests. The sensibility was assessed using the GNS-BH and GNS-BI card System (Vitek), which allows to know the clinical category and CMI value in microgram/ml. All the strains presented some resistance to antibiotics. 77.8% of the strains were resistant to more than one antibiotic and 64.9%, to more than two antibiotics. We detected 21.7% strains resistant to 4 drugs. The most frequent resistance association was ampicillin, cephoxitine, cephazoline, cephalotine, with or without amoxicillin and clavulanic acid. All the strains were sensible to cephamandol, cephuroxime, cephtriaxone, cephtazidime, cephotaxime, thicarciline, aztreonam, azlociline, mezlociline, piperaciline, gentamycin, amikacine, chloranphenicol and ciprofloxacine. All the strains were resistant to ampicillin; 60.9% of the strains were sensible to the association amoxicillin and clavulonic acid. In conclusion, the treatment of extraintestinal infections does not differs basically from the treatment of the infections caused by Gram-negative bacillus. There are a broad range of effective antibiotics: aminoglycosides (gentamycin, tobramycin, amikacine), piperacyline, cephalosporines (cephuroxime, cephotaxime, cephtazidime and cephtriaxone), chloranphenicol, penemas, monobactanes of fluorquinolone.

Three patients with beta-hemolytic group A streptococcal pneumonia were described. We emphasize the opportunistic pathogenic ability of the microorganism in the lower respiratory tract, and we describe the possible pathogenic mechanisms as well as the more frequent symptoms and complications.